An apically located hybrid guanylate cyclase-ATPase is critical for the initiation of Ca2+ signaling and motility in Toxoplasma gondii

Luning Yang, Alessandro D. Uboldi, Simona Seizova, Mary Louise Wilde, Michael J. Coffey, Nicholas J. Katris, Yoshiki Yamaryo-Botté, Martina Kocan, Ross A.D. Bathgate, Rebecca J. Stewart, Malcolm J. McConville, Philip E. Thompson, Cyrille Y. Botté, Christopher J. Tonkin

Research output: Contribution to journalArticleResearchpeer-review

2 Citations (Scopus)

Abstract

Protozoan parasites of the phylum Apicomplexa actively move through tissue to initiate and perpetuate infection. The regulation of parasite motility relies on cyclic nucleotide-dependent kinases, but how these kinases are activated remains unknown. Here, using an array of biochemical and cell biology approaches, we show that the apicomplexan parasite Toxoplasma gondii expresses a large guanylate cyclase (TgGC) protein, which contains several upstream ATPase transporter-like domains. We show that TgGC has a dynamic localization, being concentrated at the apical tip in extracellular parasites, which then relocates to a more cytosolic distribution during intracellular replication. Conditional TgGC knockdown revealed that this protein is essential for acute-stage tachyzoite growth, as TgGC-deficient parasites were defective in motility, host cell attachment, invasion, and subsequent host cell egress. We show that TgGC is critical for a rapid rise in cytosolic [Ca2+] and for secretion of microneme organelles upon stimulation with a cGMP agonist, but these deficiencies can be bypassed by direct activation of signaling by a Ca2+ ionophore. Furthermore, we found that TgGC is required for transducing changes in extracellular pH and [K] to activate cytosolic [Ca2+] flux. Together, the results of our work implicate TgGC as a putative signal transducer that activates Ca2+ signaling and motility in Toxoplasma.

Original languageEnglish
Pages (from-to)8959-8972
Number of pages14
JournalJournal of Biological Chemistry
Volume294
Issue number22
DOIs
Publication statusPublished - 31 May 2019

Keywords

  • calcium
  • Toxoplasma gondii
  • cell motility
  • cyclic GMP (cGMP)
  • cyclic nucleotide

Cite this

Yang, Luning ; Uboldi, Alessandro D. ; Seizova, Simona ; Wilde, Mary Louise ; Coffey, Michael J. ; Katris, Nicholas J. ; Yamaryo-Botté, Yoshiki ; Kocan, Martina ; Bathgate, Ross A.D. ; Stewart, Rebecca J. ; McConville, Malcolm J. ; Thompson, Philip E. ; Botté, Cyrille Y. ; Tonkin, Christopher J. / An apically located hybrid guanylate cyclase-ATPase is critical for the initiation of Ca2+ signaling and motility in Toxoplasma gondii. In: Journal of Biological Chemistry. 2019 ; Vol. 294, No. 22. pp. 8959-8972.
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title = "An apically located hybrid guanylate cyclase-ATPase is critical for the initiation of Ca2+ signaling and motility in Toxoplasma gondii",
abstract = "Protozoan parasites of the phylum Apicomplexa actively move through tissue to initiate and perpetuate infection. The regulation of parasite motility relies on cyclic nucleotide-dependent kinases, but how these kinases are activated remains unknown. Here, using an array of biochemical and cell biology approaches, we show that the apicomplexan parasite Toxoplasma gondii expresses a large guanylate cyclase (TgGC) protein, which contains several upstream ATPase transporter-like domains. We show that TgGC has a dynamic localization, being concentrated at the apical tip in extracellular parasites, which then relocates to a more cytosolic distribution during intracellular replication. Conditional TgGC knockdown revealed that this protein is essential for acute-stage tachyzoite growth, as TgGC-deficient parasites were defective in motility, host cell attachment, invasion, and subsequent host cell egress. We show that TgGC is critical for a rapid rise in cytosolic [Ca2+] and for secretion of microneme organelles upon stimulation with a cGMP agonist, but these deficiencies can be bypassed by direct activation of signaling by a Ca2+ ionophore. Furthermore, we found that TgGC is required for transducing changes in extracellular pH and [K] to activate cytosolic [Ca2+] flux. Together, the results of our work implicate TgGC as a putative signal transducer that activates Ca2+ signaling and motility in Toxoplasma.",
keywords = "calcium, Toxoplasma gondii, cell motility, cyclic GMP (cGMP), cyclic nucleotide",
author = "Luning Yang and Uboldi, {Alessandro D.} and Simona Seizova and Wilde, {Mary Louise} and Coffey, {Michael J.} and Katris, {Nicholas J.} and Yoshiki Yamaryo-Bott{\'e} and Martina Kocan and Bathgate, {Ross A.D.} and Stewart, {Rebecca J.} and McConville, {Malcolm J.} and Thompson, {Philip E.} and Bott{\'e}, {Cyrille Y.} and Tonkin, {Christopher J.}",
year = "2019",
month = "5",
day = "31",
doi = "10.1074/jbc.RA118.005491",
language = "English",
volume = "294",
pages = "8959--8972",
journal = "Journal of Biological Chemistry",
issn = "1083-351X",
publisher = "American Society for Biochemistry and Molecular Biology",
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Yang, L, Uboldi, AD, Seizova, S, Wilde, ML, Coffey, MJ, Katris, NJ, Yamaryo-Botté, Y, Kocan, M, Bathgate, RAD, Stewart, RJ, McConville, MJ, Thompson, PE, Botté, CY & Tonkin, CJ 2019, 'An apically located hybrid guanylate cyclase-ATPase is critical for the initiation of Ca2+ signaling and motility in Toxoplasma gondii', Journal of Biological Chemistry, vol. 294, no. 22, pp. 8959-8972. https://doi.org/10.1074/jbc.RA118.005491

An apically located hybrid guanylate cyclase-ATPase is critical for the initiation of Ca2+ signaling and motility in Toxoplasma gondii. / Yang, Luning; Uboldi, Alessandro D.; Seizova, Simona; Wilde, Mary Louise; Coffey, Michael J.; Katris, Nicholas J.; Yamaryo-Botté, Yoshiki; Kocan, Martina; Bathgate, Ross A.D.; Stewart, Rebecca J.; McConville, Malcolm J.; Thompson, Philip E.; Botté, Cyrille Y.; Tonkin, Christopher J.

In: Journal of Biological Chemistry, Vol. 294, No. 22, 31.05.2019, p. 8959-8972.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - An apically located hybrid guanylate cyclase-ATPase is critical for the initiation of Ca2+ signaling and motility in Toxoplasma gondii

AU - Yang, Luning

AU - Uboldi, Alessandro D.

AU - Seizova, Simona

AU - Wilde, Mary Louise

AU - Coffey, Michael J.

AU - Katris, Nicholas J.

AU - Yamaryo-Botté, Yoshiki

AU - Kocan, Martina

AU - Bathgate, Ross A.D.

AU - Stewart, Rebecca J.

AU - McConville, Malcolm J.

AU - Thompson, Philip E.

AU - Botté, Cyrille Y.

AU - Tonkin, Christopher J.

PY - 2019/5/31

Y1 - 2019/5/31

N2 - Protozoan parasites of the phylum Apicomplexa actively move through tissue to initiate and perpetuate infection. The regulation of parasite motility relies on cyclic nucleotide-dependent kinases, but how these kinases are activated remains unknown. Here, using an array of biochemical and cell biology approaches, we show that the apicomplexan parasite Toxoplasma gondii expresses a large guanylate cyclase (TgGC) protein, which contains several upstream ATPase transporter-like domains. We show that TgGC has a dynamic localization, being concentrated at the apical tip in extracellular parasites, which then relocates to a more cytosolic distribution during intracellular replication. Conditional TgGC knockdown revealed that this protein is essential for acute-stage tachyzoite growth, as TgGC-deficient parasites were defective in motility, host cell attachment, invasion, and subsequent host cell egress. We show that TgGC is critical for a rapid rise in cytosolic [Ca2+] and for secretion of microneme organelles upon stimulation with a cGMP agonist, but these deficiencies can be bypassed by direct activation of signaling by a Ca2+ ionophore. Furthermore, we found that TgGC is required for transducing changes in extracellular pH and [K] to activate cytosolic [Ca2+] flux. Together, the results of our work implicate TgGC as a putative signal transducer that activates Ca2+ signaling and motility in Toxoplasma.

AB - Protozoan parasites of the phylum Apicomplexa actively move through tissue to initiate and perpetuate infection. The regulation of parasite motility relies on cyclic nucleotide-dependent kinases, but how these kinases are activated remains unknown. Here, using an array of biochemical and cell biology approaches, we show that the apicomplexan parasite Toxoplasma gondii expresses a large guanylate cyclase (TgGC) protein, which contains several upstream ATPase transporter-like domains. We show that TgGC has a dynamic localization, being concentrated at the apical tip in extracellular parasites, which then relocates to a more cytosolic distribution during intracellular replication. Conditional TgGC knockdown revealed that this protein is essential for acute-stage tachyzoite growth, as TgGC-deficient parasites were defective in motility, host cell attachment, invasion, and subsequent host cell egress. We show that TgGC is critical for a rapid rise in cytosolic [Ca2+] and for secretion of microneme organelles upon stimulation with a cGMP agonist, but these deficiencies can be bypassed by direct activation of signaling by a Ca2+ ionophore. Furthermore, we found that TgGC is required for transducing changes in extracellular pH and [K] to activate cytosolic [Ca2+] flux. Together, the results of our work implicate TgGC as a putative signal transducer that activates Ca2+ signaling and motility in Toxoplasma.

KW - calcium

KW - Toxoplasma gondii

KW - cell motility

KW - cyclic GMP (cGMP)

KW - cyclic nucleotide

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DO - 10.1074/jbc.RA118.005491

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